HELP!sequencing

Elena Baena Gonzalez elena at phoenix.oulu.fi
Tue Dec 27 09:26:14 EST 1994


elena at phoenix.oulu.fi         at Department of Genetics. Univ. of Oulu, FIN

hello,

I'm having serious problems with sequencing. I've been using the 
Circumvent thermal cycle dideoxyDNA sequencing kit (New England 
Biolabs) for sequencing PCR products(dsDNA)of the heterozygous 
plant Cardaminopsis petraea. I get some sequence, but with so much 
background and compression that it is impossible to read it.
The protocol I'm using has been proven to work efficiently with 
bacteria, so, could all these problems be due just to the heterozy-
gosity of the template?
I have tried different primers, cycle temperatures, concentrations of
template and primer, DNA purification methods,etc.. , but without
succeed.
I would really appreciate some advice on this matter.
Thank you very much in advance.
elena at phoenix.oulu.fi
 





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